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. 2011 Dec 29;6(12):e29758. doi: 10.1371/journal.pone.0029758

Figure 1. Twist1 binds and promotes gene expression from an ECR associated with Tbx20 (Tbx20boxA).

Figure 1

A. An E-box-containing ECR is located −15040 to −14862 base pairs (bps) from the chicken Tbx20 transcriptional start site (+1). Cross-species genomic alignment of the E-box consensus sequence (shaded grey) and mutated sequence (mutated bps indicated by *) are indicated. B. Tbx20boxA or Tbx20boxA-Mut plasmids were co-transfected into HEK293 cells, with empty vector or with Twist1, E12, or Twist1 and E12 expression vectors followed by luciferase reporter assays. Fold change over the empty vector control set to 1 is shown with standard error of the mean (SEM). C. ChIP assays were performed with anti-Twist1 in mE12.5 ECCs and mE17.5 AV valves quantified by qPCR. Fold enrichment was evaluated by comparing anti-Twist1 IP of E12.5 ECCs or anti-Twist1 IP of E17.5 valves versus IgG (negative control) set to 1. Statistical significance was determined by Student's t-Test, p = ≤0.05 indicated by *. All experiments were performed in biological and technical triplicate. Histograms are a compilation of n = 3 experiments.